Monolithic column electroelution apparatus and method thereof
An electric elution and monolithic column technology, applied in the field of analytical chemistry, can solve the problems of low recovery rate, long elution time, cumbersome operation, etc., and achieve the effects of simple production, shortened elution time, and simplified operation steps
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Embodiment 1
[0033] Example 1: Electroelution of Target Proteins from Agarose Gel Electrophoresis (AGE)
[0034] Step 1: Perform protein (such as bovine serum albumin) AGE
[0035] Prepare agarose gel (0.8 % agarose, w / v) with 9 mM barbiturate buffer (pH 8.6). The sample was dissolved in 5 ml of 9 mM barbiturate buffer (pH 8.6) with standard bovine serum albumin 5 mg, and it was completely dissolved by stirring. Then add 160 microliters of sample solution to load the sample for agarose electrophoresis, the electrophoresis voltage is 100 V, and the experimental temperature is 25 o C, the electrophoresis time is 35 min. After electrophoresis, take out the gel, cut off both sides of the gel for quick staining, and put it back on the original gel plate to locate the unstained protein band after completion.
[0036] Step 2: Cut the gel plate containing the bands of the protein of interest (such as albumin) and place it in the slot
[0037] Cut according to the position of the target prote...
Embodiment 2
[0047] Example 2: Electroelution of Target Proteins from Polyacrylamide Gel Electrophoresis (PAGE)
[0048] Step 1: Perform protein (such as serum protein) PAGE
[0049] PAGE electrophoresis conditions: polyacrylamide separating gel (T: 10%, C: 2.7%), polyacrylamide stacking gel (T: 4%, C: 2.7%), 1500 mM Tris-Hcl separating gel buffer (pH 8.9 ), 500 mM Tris-Hcl stacking gel buffer (pH 6.8), 25 mM Tris-Gly electrode buffer (pH 8.3), serum samples were dissolved in 50 mM Tris-Hcl stacking gel buffer (pH 6.8, containing 1% bromine phenol blue), 10-fold dilution, 10 microliters of sample per well, electrophoresis at 60 V until the bromophenol blue enters the separation gel, and then electrophoresis at 160 V until the bromophenol blue reaches the lower end of the gel, and the electrophoresis time is 90 min. After electrophoresis, take out the gel, cut off both sides of the gel for quick staining, and put it back on the original gel plate to locate the unstained protein band afte...
Embodiment 3
[0060] Example 3: Electroelution of Target Nucleic Acids from Agarose Gel Electrophoresis (AGE)
[0061] Step 1: Perform nucleic acid AGE
[0062] Carry out DNA agarose electrophoresis according to conventional conditions, the gel concentration is 0.5%, and the gel contains EtBr 0.5 μ g / ml, the electrode buffer is TAE (40 mM, pH 8.0), and the voltage is 4V / cm. After electrophoresis, the gel was moved to the UV lamp (360 nm), and the target band was quickly cross-cut with an autoclaved sterile scalpel blade, and then the UV lamp was turned off for subsequent operations under visible light.
[0063] Step 2: Put the cut gel plate containing the nucleic acid zone into the slot
[0064] According to the target nucleic acid band drawn in step 1, remove and put the complete gel plate into the slot, the width and thickness of the gel plate match the slot, and cling to the small cylinder on the other side.
[0065] Step 3: Put the dialysis tubing on the socket and add the eluent ...
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